TPMVd Detection Kits

PDD-361

TPMVd Detection Kits

Tomato Planta Macho Viroid (TPMVd) End-Point RT-PCR Kit is designed for the detection of TPMVd specific viroid RNA based on the use of end-point RT-PCR technology.

Tomato Planta Macho Viroid(TPMVd) End-Point RT-PCR Kit is designed for the detection of TPMVd-specific RNA based on the use of end-point RT-PCR technology. This kit is designed
for research use only and not for use in diagnostic procedures. The kit includes Master Mix and primers for the specific amplification of a 248 nucleotide region of the TPMVd genome, as well as a positive control and negative control to confirm the integrity of the kit reagents. In addition, the kit contains a loading dye and a DNA ladder to facilitate the analysis of the results. The detection of TPMVd-specific RNA is based on end-point RT-PCR technology, utilizing Reverse transcription polymerase chain reaction (RT-PCR) for the amplification of specific TPMVd RNA
sequences. For analysis of the RT-PCR data, the RT-PCR reaction is loaded on an agarose RNA gel along with the provided DNA ladder for qualitative analysis.

1. Before use, suitable amounts of all components should be completely thawed at room temperature, mixed by gentle vortexing or by pipetting, and centrifuged briefly.
2. Work quickly on ice.
3. The recommended minimum number of DNA samples tested per PCR run is 6.

Viroid

1. Follow universal precautions. All specimens should be considered as potentially infectious and handled accordingly.
2. Ensure that a suitable lab coat, disposable gloves, and protective goggles are worn when handling specimens and kit reagents.
3. Use sterile pipette tips with filters. Use proper pipetting techniques and maintain the same pipetting pattern throughout the procedure to ensure optimal and reproducible values.
4. Ensure that appropriate specimen collection, transport, storage, and processing techniques are followed for optimal performance of this test.

1. MDx 2X RT-PCR Master Mix 350 µL
2. TPMVd Primer Mix 70 µL
3. TPMVd Positive Control 50 µL
4. Nuclease-Free Water 1.25 mL
5. Loading Dye 100 µL
6. DNA Ladder 100 µL
7. Product Insert 1

1. Appropriate Real-Time PCR Instrument with FAM and HEX filter channel
2. DNA Purification Kit
3. Disposable powder-free gloves
4. Benchtop microcentrifuge
5. Micropipettors
6. Sterile pipette tips with filters
7. PCR tubes
8. Vortex mixer
9. PCR reaction preparation station (Optional)

Tomato Planta Macho Viroid (TPMVd) is the causal agent of a disease of tomato known as “Planta Macho” (male plant) disease, which occurs primarily in the Mexican states of Morelos and Mexico. Symptoms of the disease resemble those induced in tomato by the potato spindle tuber viroid, but are more severe. Infected plants are severely stunted and exhibit strong epinasty of leaves and leaflets. Infected plants produce more flowers and fruits than healthy plants, however the fruit remains small and has no commercial value. In the field the disease appears to be spread via contact with diseased foliage or contaminated tools. Tomato Planta Macho Viroid RT-PCR Detection Kit is a rapid and sensitive TPMVd detection tool which can be applicable for the early diagnosis of TPMVd.

PCR

The specificity of Tomato Planta Macho Viroid (TPMVd) End-Point RT-PCR Kit is first and foremost ensured by the selection of the TPMVd-specific primers, as well as the selection of stringent reaction conditions. The TPMVd primers were checked for possible homologies to all plant viroids in GenBank published sequences by sequence comparison analysis.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.